SEC14L3 / SEC14-like protein 3 · Western blot design guide

Design a Western Blot for SEC14L3

Source-linked SEC14L3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SEC14L3 WB antibodies. Everything you need to plan the experiment before you commit precious samples.

Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use

Western blot protocol sheet for SEC14L3: expected band ~46 kDa, hero antibody A14501-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **SEC14L3** Western blot protocol sheet — expected band ~46 kDa, antibody A14501-1, controls and PMC citations. Open the full SEC14L3 WB guide →

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SEC14L3 Western Blot Experimental Design Guide

Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before running

	Expected band	~46 kDa
	Observed band	47 kDa
	Gel	5–20% (catalog A14501-1)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	—
	Caveat	—
	Gene-set association	MSigDB C7 membership
	Isoform	3 isoform(s)

Section 1

Source-Linked SEC14L3 Western Blot Protocol Options

The A14501-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.

Recommended Western blot protocol parameters
Sample / lysate	human Hek293, human U-87MG, human PC-3, rat lung, rat liver, mouse lung, mouse liver (catalog A14501-1)
Gel %	5–20% (catalog A14501-1)
Load	30ug; reducing conditions (catalog A14501-1)
Transfer	a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A14501-1)
Membrane	Nitrocellulose membrane (catalog A14501-1)
Blocking	5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A14501-1)
Primary antibody	A14501-1 · 0.5 μg/mL (catalog A14501-1)
Primary incubation	overnight at 4°C (catalog A14501-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A14501-1)
Secondary incubation	1.5 hour at RT (catalog A14501-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A14501-1)
Detection	ECL (catalog A14501-1)

Section 2

What Is the Expected SEC14L3 Western Blot Band Size?

SEC14L3 is predicted at 46 kDa and observed at approximately 47 kDa; the cause of the difference is not established.

What am I looking at on my blot?
Band at approximately 47 kDa	Empirical SEC14L3 band reported in reducing whole-cell and tissue lysates; confirm identity with controls
Band near 46 kDa	Consistent with the sequence-predicted mass, but identity requires confirmation
Single sharp band near 47 kDa	Consistent with the reported band; the listed features do not establish a visible modification shift
Two or more bands at different positions	Isoforms 1, 2, and 3 are listed, but distinct migration is unverified; confirm band identities

💡Expected SEC14L3 appearanceSEC14L3 has a predicted mass of 46 kDa and an empirical band at approximately 47 kDa; the difference is unexplained by the listed features, so confirm identity with appropriate controls.

How each factor affects band size
UniProt predicted mass	46 kDa sequence-derived mass; the empirical band is approximately 47 kDa
Isoform 1	A named splice isoform whose relative band size is not supplied
Isoform 2	A named splice isoform whose relative band size is not supplied
Isoform 3	A named splice isoform whose relative band size is not supplied

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	SEC14L3 signal may be below detection in this sample	Check loading and antibody performance with a validated positive lysate
Band higher than expected	An isoform or unrelated antibody-reactive protein is possible; no larger isoform mass is supplied	Compare with the 47 kDa reference and verify identity by SEC14L3 depletion or a second antibody
Band lower than expected	An isoform or unrelated antibody-reactive protein is possible; no smaller isoform mass is supplied	Verify identity by SEC14L3 depletion or a second antibody
Multiple bands	Three isoforms are listed, but their separate migration is unverified	Use isoform-aware validation or SEC14L3 depletion to assign bands
Weak or no signal	SEC14L3 signal may be below assay detection	Check transfer, loading, and antibody performance with a validated positive sample

Source: UniProt Q9UDX4 Human Protein Atlas product datasheet (A14501-1)

Experiment evidence

Western blot images showing SEC14L3 band

A14501-1 · SEC14L3

← swipe / scroll for more images →

Real WB data behind the band-size expectations in this section.

Sample controls for SEC14L3 Western blot

🧪For positive controls for SEC14L3 in Western blot, you can use no HPA-supported tissue or cell sample because none is reported.

Positive control: No high/medium HPA tissue identified

Negative control: Suggested KO / knockdown lysate

Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.

⚠️Feasibility: HPA provides no expression data, so tissue controls cannot be selected from this evidence.

HPA tissue expression evidence for SEC14L3

Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Higher expression tissues · candidate positive controls from IHC

Tissue	Cell type	Level	Evidence	Source
No high/medium HPA tissues identified in the supplied evidence.

Lower expression tissues · IHC evidence, not confirmed WB-negative controls

Tissue	Cell type	Level	Evidence	Source
No lower-expression tissue rows available in the supplied evidence.

Section 3

Advanced SEC14L3 Western Blot Tips

Deeper troubleshooting and optimisation questions for SEC14L3, answered from its protein features.

How should SEC14L3 band migration be interpreted?

Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.

Which SEC14L3 isoforms could produce smaller bands?

Isoforms · Isoform 2 lacks canonical residues 1–59. In isoform 3, canonical residues 1–78 are replaced by methionine. Both changes shorten the sequence, so consider these isoforms when assessing lower bands, but their presence does not establish where they migrate.

Yes, depending on its epitope. Isoform 2 lacks canonical residues 1–59, and isoform 3 replaces canonical residues 1–78. Check the antibody epitope against each sequence before interpreting a missing or weaker isoform band.

Decide whether to measure the 47 kDa band or a defined set of bands, and apply that choice consistently. Confirm that the antibody epitope is present in the isoforms you intend to quantify; the N-terminal differences could change which isoforms the antibody detects.

Do listed modifications explain a SEC14L3 band shift?

PTM · No modified residues or glycosylation sites are listed for SEC14L3. The supplied features therefore do not support assigning a band shift to either modification. An observed mass difference alone does not identify its cause.

Does this guide establish induction of SEC14L3?

Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.

How should transfer be checked for SEC14L3?

Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the A14501-1 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.

How should SEC14L3 be quantified?

Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.

Why might SEC14L3 appear at 47 kDa instead of 46 kDa?

Interpretation · The supplied observed band is 47 kDa, close to the 46 kDa predicted mass. The listed features do not establish a cause for the 1 kDa difference; use the observed position as a reference when evaluating your blot.

Consider the three listed isoforms and their N-terminal sequence differences. Compare band positions with the 47 kDa observed reference and check antibody epitope coverage. Multiple bands alone do not identify specific isoforms.

Boster reagents

SEC14L3 Western Blot Antibodies

Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.

Real WB data

Western blot analysis of SEC14L3/TAP2 using anti-SEC14L3/TAP2 antibody (A14501-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions. Lane 1: human Hek293 whole cell lysates, Lane 2: human U-87MG whole cell lysates, Lane 3: human PC-3 whole cell lysates, Lane 4: rat lung tissue lysates, Lane 5: rat liver tissue lysates, Lane 6: mouse lung tissue lysates, Lane 7: mouse liver tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SEC14L3/TAP2 antigen affinity purified polyclonal antibody (Catalog # A14501-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SEC14L3/TAP2 at approximately 47KD. The expected band size for SEC14L3/TAP2 is at 47KD.

Anti-SEC14L3/TAP2 Antibody Picoband®

Cat # A14501-1

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A14501-1 is the sole listed anti-SEC14L3 antibody. Its Western blot image shows an approximately 47 kDa band in the specified human cell, rat tissue, and mouse tissue lysates. No publication or orthogonal validation evidence was supplied.

Which to pick: Choose A14501-1; it is the only listed option. Its WB image includes HEK293, U-87MG, and PC-3 cell lysates, plus rat and mouse lung and liver lysates, using 30 µg per lane and 0.5 µg/mL primary antibody overnight at 4°C.

Source: BosterBio SEC14L3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q9UDX4.
Human Protein Atlas. SEC14L3 tissue expression.
PMC12022014 — target-verified WB comparison